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Q. Chen, W. Xiao, W. Liu, L. Li, Z. Li, D. Liang, L.D. White, D.A. Fox, P.A. Overbeek; Gene Expression Profiling In Embryonic Mouse Lenses . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2552.
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© ARVO (1962-2015); The Authors (2016-present)
At embryonic day 10 (E10), the mouse lens is a hollow sphere of epithelial cells termed the lens vesicle. By E12, the cells located at the posterior surface of the lens vesicle are induced to differentiate into the primary lens fiber cells that elongate to fill the lens vesicle to form an intact lens. Although many studies have identified genes controlling the early stages of lens formation, little is known about the global gene expression profile during this developmental transition. Using laser capture microdissection (LCM) and RNA amplification, we compared the gene expression profiles between E10 and E12 lenses.
Lens cells of C57/BL6 mouse embryos at E10 and E12 were dissected out using the PixCell II LCM System (Arcturus). Total RNA was extracted using the PicoPure RNA Isolation Kit (Arcturus) according to the manufacturer’s protocol. The RNA was amplified, labeled and hybridized to the 430 2.0 mouse chip (Affymetix) according to the manufacturer’s instructions. Data extracted from the images were analyzed using different software. Expression of the selected genes was confirmed by real–time (quantitative) PCR (Q–PCR).
Analysis of microarray data from E10 to E12 lenses identified 1690 differentially expressed genes with greater than 2–fold change in expression levels. 673 genes, including 9 subclasses of crystallin genes, were upregulated in E12 lenses compared to E10 lenses. When the differentially expressed genes were submitted to EASE (2.0) for functional group analysis, we found that 58 were classified as transcription factors, 56 showed receptor activity and 15 were G–protein coupled receptors. The gene IDs of the 1690 probe sets were submitted to GeneMerge. According to the Gene Ontology, about 5% of 673 genes are related to cell differentiation, 5% to signal transduction, 4% to G–protein coupled receptor protein signaling pathway, 10% to the regulation of transcription and 18% to cell communication. A set of genes which fit these categories was confirmed by Q–PCR.
These microarray and Q–PCR results provide new information about patterns of gene expression during early lens development. Analysis of global gene expression profiles in mouse lenses from E10 to E12 identified several previously unstudied molecular pathways that appear to be differentially regulated during very early lens development.
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